The studies reported in the present study demonstrate that Slc6a14 plays an important role in promoting the development and growth
of PyMT - driven and Neu - driven mammary tumours in mice.
We also performed microarray analysis
of PyMT - Slc6a14 + / + and PyMT - Slc6a14 − / − mammary tumours to evaluate the expression of amino acid transporters without any biased preconception of the identity of the transporters that might be subject to changes in expression in association with Slc6a14 deletion.
Not exact matches
This experiment examines the requirement
of POSTN in metastatic colonization using the MMTV -
PyMT mouse model.
The functional necessity
of POSTN was investigated by observing the pulmonary metastatic potential in POSTN - knockout MMTV -
PyMT mice, which showed a statistically significant decrease compared to controls (Malanchi et al., 2012).
b. For generation
of the MMTV -
PyMT + / tg; Postn − / − experimental female mice, male MMTV -
PyMT + / tg; Postn − / − male mice will be crossed with Postn − / − female mice.
2) The two repeated experiments analyze the source
of POSTN expression in the lung and whether it affects the number / size
of primary and secondary tumor formation in a spontaneous mouse model
of breast cancer (MMTV -
PyMT).
◯ Number
of pulmonary macrometastases in MMTV -
PyMT + / tg; Postn + / + mice relative to MMTV -
PyMT + / tg; Postn − / − mice.
Female mice carrying the MMTV -
PyMT transgene that are either Postn + / + or Postn − / − will be examined for changes in primary tumor size and the number
of spontaneously formed pulmonary macrometastases, which is a replication
of the experiment reported in Figures 3A, 3B, and Supplemental Figure 13.
Additionally, they examine MMTV -
PyMT control or POSTN null mice to test the effect
of POSTN on primary tumor growth and metastasis (Figures 3A, 3B, and S13; Malanchi et al., 2012).
c. For generation
of the MMTV -
PyMT + / tg; Postn + / + control female mice, the MMTV -
PyMT + / tg; Postn + / − male mouse will be crossed to Postn + / + female mice to obtain MMTV -
PyMT + / tg; Postn + / + male mice that will then be crossed with Postn + / + female mice.
The first experiment is to measure metastatic lung cancer in MMTV -
PyMT transgenic mice crossed with the POSTN Knock out mice, and controls, with focus on the role
of periostin in metastatic progression.
Using the MMTV -
PyMT mouse breast cancer model, which spontaneously metastasizes to the lungs, Malanchi and colleagues reported that only the CSC population, identified as CD24 + CD90 +, were capable
of initiating lung metastases and secondary metastases (Guy et al., 1992; Lin et al., 2003; Malanchi et al., 2012).
PyMT - Tg female mice typically develop tumours at the age
of ~ 10 weeks.
(A) Demonstration
of Slc6a14 mRNA up - regulation in
PyMT - driven mammary tumours in mice; N, mammary tissue from wild - type mice; T, mammary tumour tissue from
PyMT - Tg mice.
(B) Representative images
of the excised tumours and lung metastatic nodules (black arrows) from 4 - month - old Slc6a14 + / + /
PyMT mice and Slc6a14 − / − /
PyMT mice.
We then assessed the consequences
of Slc6a14 deletion on
PyMT - driven breast cancer in these mice.
However, when crossed with
PyMT - Tg mice or MMTV / Neu (mouse mammary tumour virus promoter - Neu)- Tg mice, the development and progression
of breast cancer were markedly decreased on Slc6a14 − / − background.
With 12 different amino acid transporters examined by RT - PCR, none
of the transporters differed in expression between
PyMT - driven tumours with Slc6a14 + / + and Slc6a14 − / − backgrounds (result not shown).
We examined the expression
of Slc6a14 in breast tumours obtained from 4 - month - old
PyMT - Tg female mice and in mammary glands obtained from age - matched wild - type female mice.
PyMT - Tg mice develop breast cancer spontaneously at ~ 10 weeks
of age [19].
At 3 months
of age, all Slc6a14 + / + females with
PyMT transgene developed tumours in multiple mammary glands which grew to sizes
of 2000 — 2500 mm3 at 4 months
of age (Figure 2B).
In contrast, no Slc6a14 − / − female with
PyMT transgene developed tumours at 3 months
of age; however, at 4 months
of age, small tumour nodules became detectable (Figure 2B), indicating a marked delay in mammary tumour development in
PyMT - Tg mice as a consequence
of Slc6a14 deletion.
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